Microfluidic matrix localization apparatus and methods

Abstract

Multiphasic microfluidic apparatus for performing product fluid manipulation and separation in a single continuous unit are provided. Related methods, kits, and compositions are also provided.

What is claimed is:  
     
       1. A method of performing a polymerase chain reaction in a microfluidic apparatus, said method comprising: 
       providing a microfluidic device comprising at least two intersecting channels in fluid communication;  
       filling at least one of said two intersecting channels with an electrophoretic phase selected from the group consisting of a sieving matrix and a molecular partition matrix, wherein the electrophoretic phase comprise at least two PCR reactants and a thermostable polymerase dispersed therein;  
       performing the polymerase chain reaction in the presence of the electrophoretic phase to produce at least a first product.  
     
     
       2. The method of claim  1 , said method further comprising a step of separating the at least first product from the reactants in the presence of the electrophoretic phase. 
     
     
       3. The method of claim  1 , wherein at least one of said intersecting channels comprise a reaction region. 
     
     
       4. The method of claim  1 , wherein the microfluidic device comprises a high-resistance channel region for heating the at least two PCR reactants. 
     
     
       5. The method of claim  1 , wherein the two PCR reactants are heterogeneously dispersed throughout at least a portion of the electrophoretic phase. 
     
     
       6. The method of claim  1 , wherein the two PCR reactants are homogeneously dispersed throughout at least a portion of the electrophoretic phase. 
     
     
       7. The method of claim  1 , wherein at least one of the two PCR reactants is heterogeneously dispersed in at least a portion of the electrophoretic phase and one of the two PCR reactants is homogeneously dispersed throughout the electrophoretic phase. 
     
     
       8. A method of performing a polymerase chain reaction in a microfluidic apparatus, said method comprising: 
       providing a microfluidic device comprising at least two intersecting channels in fluid communication;  
       filling at least one of said two intersecting channels with a mixture comprising a sieving matrix, a thermostable polymerase and a plurality of PCR reaction components;  
       performing the PCR in the presence of the sieving matrix to produce at least a first product.  
     
     
       9. The method of claim  8 , said method further comprising a step of separating the at least first product from the PCR reaction components in the presence of the sieving matrix. 
     
     
       10. The method of claim  8 , wherein the step of separating is carried out in a channel region different from a channel region for performing the PCR. 
     
     
       11. The method of claim  8 , wherein the sieving matrix is selected from a group consisting of agarose, linear polyacrylamide, methylcellulose, polyethylene oxide and hydroxy ethyl cellulose. 
     
     
       12. The method of claim  8 , wherein one of said at least two intersecting channels have an interior dimension of between about 0.1 μm and 500 μm. 
     
     
       13. The method of claim  8 , wherein the two PCR reactants are heterogeneously dispersed throughout at least a portion of the sieving matrix. 
     
     
       14. The method of claim  8 , wherein the two PCR reactants are homogeneously dispersed throughout at least a portion of the sieving matrix. 
     
     
       15. The method of claim  8 , wherein at least one of the two PCR reactants is heterogeneously dispersed in at least a portion of the sieving matrix and one of the two PCR reactants is homogeneously dispersed throughout the sieving matrix. 
     
     
       16. The method of claim  8 , wherein one of said at least two intersecting channels comprise a reaction region. 
     
     
       17. The method of claim  16 , wherein the reaction region is substantially filled with the sieving matrix. 
     
     
       18. The method of claim  8 , wherein one of said at least two intersecting channels comprise a separation region. 
     
     
       19. The method of claim  18 , wherein the separation region is substantially filled with the sieving matrix.